This invention relates to methods and novel compounds for treating autoimmune and vascular disease. In particular, the invention relates to the use of lipophosphoglycan and novel lipophosphoglycan analogs to treat these diseases.
Pro-inflammatory cytokines, such as interleukin-1 (IL-1), tumor necrosis factor (TNF-xcex1), and interleukin-6 (IL-6) are important mediators of sepsis, rheumatoid arthritis and reactive arthritis. They are also associated with other conditions such as atherosclerosis, progression of disease in persons infected with the human immunodeficiency virus, and autoimmune disorders. There were 250,000 reported cases of sepsis in the United States in 1987. Rheumatoid arthritis in the adult population is estimated at 1%, with an additional 1% of the population estimated to have reactive arthritis or juvenile rheumatoid arthritis. Therefore, pro-inflammatory cytokines play a direct role in the pathogenesis of conditions in nearly 3 million persons. Current therapeutic methods are not very effective in reducing the morbidity and/or mortality associated with these diseases.
Depressed cellular immunity and IL-1xcex2 production are associated with leishmaniasis caused by Leishmania donovani. Leishmania donovani is an obligate intracellular parasite of the mammalian macrophage. It is worldwide in distribution and annually causes disease in 20 million persons. The life cycle of Leishmania parasites consists of two stages. The flagellated promastigote form propagates in the alimentary tract of its insect vector (the sandfly) and develops into the infectious metacyclic promastigote stage. During the sandfly""s blood meal, the metacyclic promastigotes are inoculated into the mammalian host. The promastigotes evade local host defense, disseminate, and enter macrophages throughout the reticuloendothelial system. Within macrophages they transform into the amastigote form which remains for the life of the new host. S. J. Turco and A. Descoteaux, xe2x80x9cThe Lipophosphoglycan of Leishmania parasites,xe2x80x9d Annu. Rev. Microbiol., 46:65-94 (1992); J. L. Ho, R. Badaro, D. Hatzigeorgiou, S. G. Reed, and W. D. Johnson, Jr., xe2x80x9cCytokines in the Treatment of Leishmaniasis: From Studies of Immunopathology to Patient Therapy,xe2x80x9d Biother. 7:223-35 (1994).
The two poles of immune responses in patients infected with L. donovani are: 1) a delayed type hypersensitivity (xe2x80x9cDTHxe2x80x9d) and CMI to Leishmania antigens which is associated with containment of infection, and 2) B cell activation with absence of DTH, and T-cell reactivity that is associated with visceral leishmaniasis (xe2x80x9cVLxe2x80x9d). J. L. Ho, R. Badaro, D. Hatzigeorgiou, S. G. Reed, and W. D. Johnson, Jr., xe2x80x9cCytokines in the Treatment of Leishmaniasis: From Studies of Immunopathology to Patient Therapy,xe2x80x9d Biother. 7:223-35 (1994). Patients with acute VL have both a functional T-cell defect with decreased production of IL-2 and IFN-xcex3 in response to Leishmania antigens or mitogens, and a presumed monocytic defeat with decreased in vitro production of IL-1xcex2 and TNF-xcex1 in response to Leishmania systate, LPS, or Listeria. E. M. Carvalho, R. Badaro, S. Reed, W. D. Johnson, Jr., and T. Jones, xe2x80x9cAbsence of IFN-xcex3 and IL-2 Production During Active Visceral Leishmaniasis,xe2x80x9d J. Clin. Invest, 76:2066-9 (1985). These defects are attributed either to Leishmania products, or to a predominance of the T helper-2 (xe2x80x9cTh2xe2x80x9d) type of lymphocytes. S. J. Turco and A. Descoteaux, xe2x80x9cThe Lipophosphoglycan of Leishmania Parasites,xe2x80x9d Annu. Rev. Microbiol., 46:65-94 (1992); P. Scott, xe2x80x9cIFN-xcex3 Modulates the Early Development of Th1 and Th2 Responses in a Murine Model of Cutaneous Leishmaniasis,xe2x80x9d J. Immunmol., 147:3149-3155 (1991); K. Varkla, R. Chatelain, L. M. C. C. Leal, and L. Coffman, xe2x80x9cReconstitution of C.B-17 SCID Mice with BALB/c T cells Initiates a T Helper Type-1 Response and Renders Them Capable of Healing Leishmania Major Infection,xe2x80x9d Eur. J. Immunol. 23:262-268 (1992); C. L. Karp, S. H. El-Sagi, T. A. Wynn, M. M. H. Satti, H. M. Kordojani, F. A. Hashim, M. Hag-Ali, F. A. Neva, T. B. Nutman, and D. L. Sacks, xe2x80x9cIn vivo Cytokine Profiles in Patients with Kala-azar: Marked Elevation of Both Interleukin 10 and Interferon-gamma,xe2x80x9d J. Clin. Invest. 91:1644-8 (1993); H. G. Ghalib, M. R. Piuvezam, Y. A. W. Skeiky, M. Siddig, F. A. Hashim, A. M. El-Hassan, D. M. Russo, and S. G. Reed, xe2x80x9cInterleukin 10 Production Correlates with Pathology in Human Leishmania donovani Infections,xe2x80x9d J. Clin. Invest. 92:324-9 (1993); L. Morris, A. B. Troutt, E. Handman, and A. Kelso, xe2x80x9cChanges in the Precursor Frequencies of IL-4 and IFN-xcex3 Secreting CD4+ Cells Correlate with Resolution of Lesions Inmurin Cutaneous Leishmaniasis,xe2x80x9d J. Immunol. 149:2715-2721 (1992); M. D. Sadick, F. P. Heinzel, B. J. Hodaday, R. T. Pu, R. S. Dawkins, and R. M. Locksley, xe2x80x9cCure of Murine Leishmaniasis with Anti-interleukin 4 Monoclonal Antibody; Evidence for a T cell-Dependent, Interferon-xcex3-independent Mechanism,xe2x80x9d J. Exp. Med. 171:115-127 (1990); L. M. Leal, D. W. Moss, R. Kuhn, W. Muller, and F. Y. Liew, xe2x80x9cInterleukin-4 Transgenic Mice of Resistant Background are Susceptible to Leishmania Major Infection,xe2x80x9d Eur. J. Immunol. 23:566-5690 (1993).
One unique characteristic of Leishmania infection is that the macrophage is unable to kill the parasite. In macrophages infected with Leishmania or treated with lipophosphoglycan (LPG) of promastigotes of L. donovani, impaired macrophage functions including production of IL-1xcex2 have been reported. S. J. Turco and A. Descoteaux, xe2x80x9cThe Lipophosphoglycan of Leishmania Parasites,xe2x80x9d Annu. Rev. Microbiol. 46:65-94 (1992); T. B. McNeely and S. Turco, xe2x80x9cRequirements of Lipophosphoglycan for Intracellular Survival of Leishmania donovani Within Human Monocytes,xe2x80x9d J. Immunol. 144:2745-50 (1992); S. Frankenburg, V. Leibovici, N. Mansbach, S. J. Turco, and G. Rosen, xe2x80x9cEffect of Glycolipids of Leishmania Parasites on Human Monocyte Activity,xe2x80x9d J. Immunol. 145:4284-89 (1990); S. Frankenburg, A. Gross, and V. Leibovici, xe2x80x9cLeishmania major and Leishmania donovani: Effect of LPG-containing and LPG-deficient Strains on Monocyte Chemotaxis and Chemiluminescence,xe2x80x9d Exp. Parsitol. 75:442-8 (1992); A. Descoteaux and G. Matlashewski, xe2x80x9cC-fos and Tumor Necrosis Factor Gene Expression in Leishmania donovani-infected Macrophages,xe2x80x9d Molecular Cell Biol. 9:5223-7 (1989); N. E. Reiner, W. Ng, C. B. Wilson, R. McMaster, and S. K. Burchett, xe2x80x9cModulation of in vitro Monocyte Cytokine Response to Leishmania donovani. Interferon-xcex3 Prevents Parasite-induced Inhibition of Interleukin-1 Production and Primes Monocytes to Respond to Leishmania by Producing Both Tumor Necrosis Factor-xcex1 and Interleukin-1,xe2x80x9d J. Clin. Invest. 85:1914-240 (1990). Similarly, diminished macrophage activation and/or IL-1 production have been reported to Th-2 derived cytokines such as, IL-4, IL-6, and IL-10. Essner, R. K. Rhoades, W. H. Mcbride, D. L. Morton, and J. S. Economou, xe2x80x9cIL-4 Down-regulates IL-1 and TNF Gene Expression in Human Monocytes,xe2x80x9d J. Immunol. 142:3857-61 (1989); J. L. Ho, S. H. He, M. J. C. Rios, and E. A. Wick, xe2x80x9cInterkeukin-4 Inhibits Human Macrophage Activation by Tumor Necrosis Factor, Granulocyte-macrophage Colony Stimulating Factor and Interleukin-3 for Anti-leishmania Activity and Oxidative Burst Capacity,xe2x80x9d J. Infect. Dis. 165.344-51 (1992); D. H., Hatzigeorgiou, S. H. He, J. Sobel, A. Hafner, K. Grabstein, and J. L. Ho, xe2x80x9cInterleukin-6 Down-modulates Cytokine-enhanced Antileishmanial Killing,xe2x80x9d J. Immunol. 151:3682-92 (1993); C. Bogdan, J. Paik, Y. Vodovotz, and C. Nathan, xe2x80x9cContrasting Mechanisms for Suppression of Macrophage Cytokine Release by Transforming Growth Factor-xcex3 and Interleukin-10,xe2x80x9d J. Biol. Chem. 267:23301-8 (1992). IL-1xcex2 produced by macrophages and other antigen-presenting cells mediates T-cell activation and proliferation and triggers T-cell production of IL-2. M. Luqman, L. Greenbaum, D. Lu, and K. Bottomly, xe2x80x9cDifferential Effect of Interleukin 1 on Naive and Memory CD4+ T Cells,xe2x80x9d Eur. J. Immunol. 22:95-100 (1992); P. H. Stein and A. Singer, xe2x80x9cSimilar Co-stimulation Requirements of CD4+ and CD8+ Primary T Helper Cells: Role of IL-1 and IL-6 in Inducing IL-2 Secretion and Subsequent Proliferation,xe2x80x9d Int. Immunol. 3:327-35 (1992). In addition, IL-1 plays an obligate role in the in vivo induction of activated macrophages against intracellular Listeria and activates anti-Leishmania activity in vitro,xe2x80x9d H. W. Rogers, K. C. F. Sheehan, L. M. Brunt, S. K. Dower, E. R. Unanue, and R. D. Schreiber, xe2x80x9cInterleukin 1 Participates in the Development of Anti-Listeria Response in Normal and SCID Mice,xe2x80x9d Proc. Natl. Acad. Sci. USA. 89:1011-15 (1992); D. Hatzigeorgiou, S. G. Reed, and W. D. Johnson, Jr., xe2x80x9cCytokines in the Treatment of Leishmaniasis: From Studies of Immunopathology to Patient Therapy,xe2x80x9d Biother. 7:223-35 (1994), which are hereby incorporated by reference. Therefore, inhibition of IL-1xcex2 production may be an important mechanism for evasion of CMI.
LPG, one of the most abundant surface molecules of the promastigotes but not synthesized by the amastigotes of L. donovani, plays a critical role for intramacrophage survival of promastigotes. S. J. Turco and A. Descoteaux, xe2x80x9cThe Lipophosphoglycan of Leishmania Parasites,xe2x80x9d Annu. Rev. Microbiol. 46:65-94 (1992). Although the production of IL-1xcex2 in response to LPS in human macrophages treated with LPG has been shown to be depressed, the molecular mechanisms remain undefined. S. Frankenburg, V. Leibovici, N. Mansbach, S. J. Turco, and G. Rosen, xe2x80x9cEffect of Glycolipids of Leishmania Parasites on Human Monocyte Activity,xe2x80x9d J. Immunol. 145:4284-9 (1990). Furthermore, the mechanisms reported for L. donovani-maastigote-infected human macrophages differ from those reported for murine macrophages. N. E. Reiner, xe2x80x9cParasite Accessory Cell Interaction in Murine Leishmaniasis. Evasion and Stimulus-dependent Suppression of the Macrophage Interleukin 1 Response by Leishmania donovani,xe2x80x9d J. Immunol. 138:1919-25 (1987); N. E. Reiner, W. Ng, and W. R. McMaster, xe2x80x9cParasite-accessory Cell Interactions in Murine Leishmaniasis. II. Leishmania donovani Suppress Macrophage Expression of Class I and Class II Major Histocompatibility Gene Products,xe2x80x9d J. Immunol. 138:1926-32 (1987); N. E. Reiner, W. Ng, C. B. Wilson, R. McMaster, and S. K. Burchett, xe2x80x9cModulation of in vitro Monocyte Cytokine Response to Leishmania donovani. Interferon-xcex3 Prevents Parasite-induced Inhibition of Interleukin-1 Production and Primes Monocytes to Respond to Leishmania by Producing Both Tumor Necrosis Factor-xcex1 and Interleukin-1,xe2x80x9d J. Clin. Invest. 85:1914-240 (1990).
The present invention relates to a method of treating inflammatory diseases in mammals. This process involves administering an effective amount of lipophosphoglycan or a lipophosphoglycan analogue to the mammal.
Another aspect of the present invention relates to a method of inhibiting production of adhesion molecules on endothelial cells. This process involves administering an effective amount of lipophosphoglycan to the endothelial cells.
The present invention also relates to inhibiting production of tissue factor by endothelial cells. This process involves administering an effective amount of lipophosphoglycan or lipophosphoglycan analogue to the endothelial cells.
A method of inhibiting induction of nitric oxide synthase by macrophages is also disclosed. This involves administering an effective amount of lipophosphoglycan or lipophosphoglycan analogue to macrophages.
A method of reversing the inhibitory effects of lipophosphoglycan on endothelial cells or macrophages is also disclosed. Here, an effective amount of lipophosphoglycan analogues are administered to endothelial cells or macrophages which are being contacted with lipophosphoglycan.
The present invention also relates to a method of targeting material to endothelial cells, fibroblasts, or monocytes. This involves administering lipophosphoglycan or lipophosphoglycan analogues to endothelial cells, fibroblasts, or monocytes. A product for such targeting is also disclosed.
Another aspect of the present invention relates to an isolated DNA molecule suitable for connection to a gene capable of transcription. Lipophosphoglycan or lipophosphoglycan analogues bind to the DNA molecule or trigger nuclear protein(s) binding to the DNA molecule.
Such binding antagonizes transcription of the gene. The DNA molecule, when positioned to regulate transcription of the gene, can be used to carry out a method of testing drugs for inhibiting production of tissue factor, nitric oxide synthase, proinflammatory cytokines, or adhesion molecules.